Qing Yu-ling, Zhao Jia-jiang, Ren Hong
Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Zhonghua Gan Zang Bing Za Zhi. 2004 Apr;12(4):201-4.
To investigate whether GM-CSF can enhance the antiviral effect of HBsAg DNA vaccine.
Divided animals into 8 groups. Group A: pcDNA3.1-S 100microg; Group B: pcDNA3.1-GM-CSF-S 100microg; Group C: pcDNA3.1-S-GM-CSF 100microg; Group D: pcDNA3.1-S 50microg + pcDNA3.1-GM-CSF 50microg; Group E: pcDNA3.1-GM-CSF 100microg; Group F: recombinant HBsAg vaccine 1microg; Group G: pcDNA3.1,100microg; Group H: PBS 100microl. Serum HBsAg level and concentration of IL-2, IL-4 and IFN-gamma were examined using commercial ELISA kit. The [3H] thymidine incorporation into DNA of Spleen cells was measured; HBsAg expression of hepatocytes from HBV-transgenic mice was assessed using immunohistochemical analysis.
Serum HBsAg level was lower and concentration of IL-2, IFN-gamma and SI was higher in mice immunized with pcDNA3.1-GM-CSF-S than those from mice immunized with pcDNA3.1-S and other groups (F=11.262, P<0.01, F=8.147, P<0.01, F=62.275, P<0.01, F=116.28, P<0.01. Less Hepatic HBsAg expression and decline of pcDNA3.1-GM-CSF-S of mice immunized with pcDNA3 were observed in comparison with control groups (F=41.439, P<0.01). Liver histological analysis showed no evidence of necrosis or inflammation in various groups.
The plasmid co expressing GM-CSF and HBsAg could improve HBsAg-specific humoral and cellular immune responses induced by plasmid encoding HBsAg alone and enhance HBsAg DNA vaccine antivirus effect.
研究粒细胞-巨噬细胞集落刺激因子(GM-CSF)是否能增强乙肝表面抗原(HBsAg)DNA疫苗的抗病毒效果。
将动物分为8组。A组:pcDNA3.1-S 100微克;B组:pcDNA3.1-GM-CSF-S 100微克;C组:pcDNA3.1-S-GM-CSF 100微克;D组:pcDNA3.1-S 50微克 + pcDNA3.1-GM-CSF 50微克;E组:pcDNA3.1-GM-CSF 100微克;F组:重组HBsAg疫苗1微克;G组:pcDNA3.1,100微克;H组:PBS 100微升。使用商用酶联免疫吸附测定(ELISA)试剂盒检测血清HBsAg水平以及白细胞介素-2(IL-2)、白细胞介素-4(IL-4)和γ-干扰素(IFN-γ)的浓度。测定[3H]胸腺嘧啶核苷掺入脾细胞DNA的情况;采用免疫组织化学分析评估乙肝病毒(HBV)转基因小鼠肝细胞的HBsAg表达。
用pcDNA3.1-GM-CSF-S免疫的小鼠血清HBsAg水平低于用pcDNA3.1-S免疫的小鼠及其他组,IL-2、IFN-γ和SI的浓度高于用pcDNA3.1-S免疫的小鼠及其他组(F = 11.262,P < 0.01;F = 8.147,P < 0.01;F = 62.275,P < 0.01;F = 116.28,P < 0.01)。与对照组相比,用pcDNA3.1-GM-CSF-S免疫的小鼠肝组织中HBsAg表达减少且pcDNA3.1-GM-CSF-S下降(F = 41.439,P < 0.01)。肝脏组织学分析显示各实验组均无坏死或炎症迹象。
共表达GM-CSF和HBsAg的质粒能增强单独编码HBsAg的质粒诱导的HBsAg特异性体液免疫和细胞免疫反应,并增强HBsAg DNA疫苗的抗病毒效果。